*Background/Disease Information: C. perfringens is a bacteria that leads to necrosis, bacteremia and gas gangrene. It is the most common bacterial reagent leading to gas gangrene. It is the third most common cause of food poisoning in the United Kingdom and the United States.
-- PDB # or closest PDB entry if using homology model: 2WXT
Druggable Target (see Databases for this):
Inhibitors
Inhibitors
Commentary
Organism
Structure
EDTA
enzyme activity against 1-myristoyl-lysophosphatidylcholine, glycerophosphorylcholine, and p-nitrophenyl phosphorylcholine are readily inhibited by EDTA. Indicates that a divalent cation is essential for catalytic activity
enzyme activity against 1-myristoyl-lysophosphatidylcholine, glycerophosphorylcholine, and p-nitrophenyl phosphorylcholine are readily inhibited by EGTA. Indicates that a divalent cation is essential for catalytic activity
the catalytic activity of NPP6 toward p-nitrophenyl phosphorylcholine is partially inhibited by the divalent cations. Attempts are unsuccessful to recover the activity of EDTA treated NPP6 by adding various divalent cations
Homo sapiens
-
Expression Information (has it been expressed in bacterial cells): homo sapiens, E. Coli and mus musculus. Yes it was expressed in E. Coli
Purification Method:
mono Q ion exchange chromatography column and eluted with a linear gradient of NaCl (0-2 M) using the deltaKTA system in homo sapiens and mus musculus
*length of your protein in Amino Acids 474 Molecular Weight of your protein 53611.1 Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water 77935
*NCBI Gene # or RefSeq#: NCTC8237
*Protein ID (NP or XP #) or Wolbachia#:GenBank: CAA31943.1
http://www.ncbi.nlm.nih.gov/protein/CAA31943.1
>gi|194293816|gb|EU839779.1| Clostridium perfringens strain S01 phospholipase C (plc) gene, complete cds
*Organism: Clostridium Perfrigens
*Background/Disease Information: C. perfringens is a bacteria that leads to necrosis, bacteremia and gas gangrene. It is the most common bacterial reagent leading to gas gangrene. It is the third most common cause of food poisoning in the United Kingdom and the United States.
ESSENTIALITY
Molecular cloning and nucleotide sequence of the alpha-toxin (phospholipase C) of Clostridium perfringens.
http://iai.asm.org/content/57/2/367.short
Complex of enzymes:
*EC#: 3.1.4.3
Link to BRENDA EC# page: http://brenda-enzymes.org/literature/lit.php4?e=3.1.4.3&r=665632
--Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Enzyme_Assay/phospholipaseccper.Par.0001.File.tmp/phospholipaseccper.pdf
--
Structure Available (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 2WXT
Druggable Target (see Databases for this):
Inhibitors
Expression Information (has it been expressed in bacterial cells): homo sapiens, E. Coli and mus musculus. Yes it was expressed in E. Coli
Purification Method:
mono Q ion exchange chromatography column and eluted with a linear gradient of NaCl (0-2 M) using the deltaKTA system in homo sapiens and mus musculus
*Amino Acid Sequence: *
1 MSRLVVVSNRIAPPDEHAASAGGLAVGILGALKAAGGLWFGWSGETGNEDQPLKKVKKGN
61 ITWASFNLSEQDLDEYYNQFSNAVLWPAFHYRLDLVQFQRPAWDGYLRVNALLADKLLPL
121 LQDDDIIWIHDYHLLPFAHELRKRGVNNRIGFFLHIPFPTPEIFNALPTYDTLLEQLCDY
181 DLLGFQTENDRLAFLDCLSNLTRVTTRSAKSHTAWGKAFRTEVYPIGIEPKEIAKQAAGP
241 LPPKLAQLKAELKNVQNIFSVERLDYSKGLPERFLAYEALLEKYPQHHGKIRYTQIAPTS
301 RGDVQAYQDIRHQLENEAGRINGKYGQLGWTPLYYLNQHFDRKLLMKIFRYSDVGLVTPL
361 RDGMNLVAKEYVAAQDPANPGVLVLSQFAGAANELTSALIVNPYDRDEVAAALDRALTMS
421 LAERISRHAEMLDVIVKNDINHWQECFISDLKQIVPRSAESQQRDKVATFPKLA
*length of your protein in Amino Acids 474
Molecular Weight of your protein 53611.1
Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water 77935
*GC% Content for gene: 34%
Primer design
1 ATGTCTCGTCTCGTTGTTGTTTCTAATCGTATCGCGCCTCCAGACGAACACGCGGCGTC 59
2 GCCGCCTTGAGGGCACCGAGGATACCCACCGCGAGGCCACCAGCAGACGCCGCGTGTTCG 60
3 TGCCCTCAAGGCGGCAGGCGGTCTGTGGTTCGGTTGGTCCGGTGAGACCGGTAACGAGGA 60
4 CCCAGGTGATGTTACCTTTCTTAACTTTTTTGAGTGGCTGATCCTCGTTACCGGTCTCAC 60
5 AGAAAGGTAACATCACCTGGGCGTCTTTCAACCTGTCTGAACAAGACCTGGACGAATACT 60
6 GGAACGCAGGCCAGAGAACGGCGTTAGAGAACTGGTTGTAGTATTCGTCCAGGTCTTGTT 60
7 TCTCTGGCCTGCGTTCCATTACCGTCTCGACCTGGTGCAATTTCAGCGTCCAGCGTGGGA 60
8 GGGAGGAGTTTATCCGCCAGCAGCGCGTTAACACGCAGATAGCCGTCCCACGCTGGACGC 60
9 TGGCGGATAAACTCCTCCCGCTCCTCCAGGACGACGATATCATCTGGATTCACGACTACC 60
10 CGCCACGCTTACGCAGCTCGTGCGCGAACGGCAGGAGGTGGTAGTCGTGAATCCAGATGA 60
11 CTGCGTAAGCGTGGCGTAAATAACCGTATCGGTTTCTTCCTGCACATCCCGTTCCCAACC 60
12 CAGCAGGGTGTCGTAGGTCGGGAGTGCGTTAAAGATTTCCGGGGTTGGGAACGGGATGTG 60
13 ACCTACGACACCCTGCTGGAACAGCTCTGTGACTACGACCTGCTCGGCTTCCAGACCGAG 60
14 GGGTCAGATTAGAGAGGCAGTCCAGAAACGCCAGACGGTCATTCTCGGTCTGGAAGCCGA 60
15 CTGCCTCTCTAATCTGACCCGTGTTACCACTCGTAGCGCGAAATCTCATACTGCGTGGGG 60
16 TTCGATACCGATAGGGTAAACTTCGGTGCGGAACGCTTTACCCCACGCAGTATGAGATTT 60
17 AAGTTTACCCTATCGGTATCGAACCGAAAGAAATCGCGAAACAAGCAGCCGGTCCGCTGC 60
18 TTCTGAACGTTTTTCAGTTCCGCTTTGAGCTGGGCCAGCTTTGGTGGCAGCGGACCGGCT 60
19 CGGAACTGAAAAACGTTCAGAACATCTTCTCTGTTGAACGCCTGGACTACTCTAAAGGTC 60
20 TTTCCAGGAGCGCTTCGTACGCCAGGAAACGCTCCGGCAGACCTTTAGAGTAGTCCAGGC 60
21 ACGAAGCGCTCCTGGAAAAATACCCGCAACATCATGGTAAAATTCGTTACACCCAGATTG 60
22 CTTGGTACGCCTGCACGTCGCCACGAGAGGTCGGCGCAATCTGGGTGTAACGAATTTTAC 60
23 CGTGCAGGCGTACCAAGACATCCGTCACCAACTGGAGAACGAAGCAGGCCGCATCAACGG 60
24 GGTTCAGGTAGTACAGCGGAGTCCAACCCAGCTGACCGTACTTACCGTTGATGCGGCCTG 60
25 CCGCTGTACTACCTGAACCAACACTTCGACCGTAAACTGCTGATGAAAATCTTCCGTTAC 60
26 TACCGTCGCGCAGTGGGGTCACCAGACCAACGTCAGAGTAACGGAAGATTTTCATCAGCA 60